Application
Western blot
Loading amount
80 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient gel)
Sample
Mouse Cell lysate - whole cell (NIH3T3)
Specification
NIH3T3
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/500
Secondary antibody
Name
Non-Abcam antibody was used: HRP-goat-anti-rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL
Bands
Specific: 60, 110 kDa Non-specific: >150, 40, 20 kDa
Exposure
2 minute(s) and 0 second(s)
Additional data
Abcam response
The reviewer requested suggestions for other antibodies that are expected to react with most if not all PDE4 isoforms. We suggest ab14628 as an alternative but a mixture of antibodies may be the best approach.
We also suggested denaturing the samples at a lower temperature, for instance 60C, to minimize what appears to be aggregation of PDE4B at high molecular weights.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Xuecai Ge
Verified customer
投稿 Apr 03 2014